Chitosan Nanocarrier Study

We made Chitosan nanospheres for drug delivery applications,Using a water in oil emulsion method followed by glutaraldehyde crosslinking of the chitosan amino groups ,Use trypsin for tcontained a terminal amine , glutaraldehyde addition indiscriminately bound the active agent to the polymer as it did between chitosan chains,coated with carboxymethyl chitosan ,causing drug immobilization rather than encapsulation. New methods of Chloroacetic acid - lsopropylalcohol was utilised for preparation of carboxymethyl chitosan, reclaim by 87.8％, new methods of chlorosulfonic acid - pyridine for sulphate chitosan , reclaim by 110.67％ , new methods of Ethylene Oxide for preparation of hydroxyl chitosan, reclaim by 87％, FTIR spectra indicate the substituent numbers and positions. column chromatography analysis the molecular weight and distribution.Chitosan nanospheres made by mixers of chitosan and derivative of chitosan at 3:1 with emulsification process and microspheres enveloped with carboxymethyl chitosan.chitosan/carboxymethyl chitosan nanospheres, chitosan/sulphate chitosan nanospheres,chitosan/hydroxyl chitosan nanospheres, reclaim by 75.33%,79%,79.67%,trypsin loaded by 29.3%,24.7%,22.2%., FTIR can detect the carboxymethyl chitosan cover. Chitosan core-coated microspheres were morphologically characterized for shape, surface characteristics and size distribution , nanospheres were determined with Scanning Electron Microscope that uniformly distribution from 10-100 μm . This method carried out in mild conditions and was particularly useful for miscroencap solution of thermally sensitive proteins. the collapse time constituted by different ramification of chitosan and chitosan ratios(optimal balue was 1:3) .and Glutaraldehyde at different concentrations can modulate drug releasem(optimal balue was 0.4%),the release of trypsin sustain for 24 hour.chitosan/carboxymethyl chitosan nanospheres Km =4.20x10-3mol/L, chitosan/sulphate chitosan nanospheres Km =3.87x10-3mol/L , chitosan/hydroxyl chitosan nanospheres, Km =3.92x10-3mol/L, compared to origin trypsin Km =3.40x10-3mol/L,that indicated the activety of binding to substrate not have significant difference .Trypsin contents on chitosan microspheres have more stabilize on pH 6-10, optimal at pH 8.0 , immobilized trypsin had high activity at temperature as 80℃ and after 80% alcohol denaturants still high activity. Trypsin release profiles were also investigated, Use Trypsin microspheres to digest HeLa cell ,after 7day culture, None pollution detected, that indicated the methed of immobiliztion was a aseptic technique. Cell curve and MTT indicate that Chitosan nanospheres have no cell toxicity.